Avaliação do potencial antiinflamatório, anticoagulante e hemorrágico de heparinóides presentes em "Artemia franciscana"

Heparan sulfate (HS) and Heparin (Hep) glycosaminoglycans (GAGs) are heterogeneous and highly charged polysaccharides. HS is structurally related to Hep but is much less substituted with sulfo groups than heparin and has a more varied structure (or sequence). Because of structural similiarities between these two polymers, they have been described together as heparinoids . Both chains bind a variety of proteins and mediate various physiologically important processes including, blood coagulation, cell adhesion and growth factor regulation. Heparinoids with structural characteristics similar to these described from HS and/or Hep from mammalian tissues have been isolated from different species of invertebrates, although only a few heparinoids from unusual sources have been characterized. The present study describes the presence of unusual heparinoids population from Artemia franciscana, isolated after proteolysis and fractionation by ion exchange resin and named, F-3.0M. The study model in vivo were hemostasis (rat tail scarification) and inflamatoty activity. The tests in vitro were used for coagulations assays (PT and APTT). The analyse of the heparinoids eluted with 3,0M NaCl showed electrophoretic migration in different buffer systems a single band with a behaviour intermediate between those of mammalian HEP and HS. The main products obtained from Artemia heparinoids after enzymatic degradation with heparitinases I and II from F. heparinum were N-sulphated disaccharides (∆U-GlcNS,6S/ ∆U,2S-GlcNS and ∆U-GlcNS) and N-acetylated disaccharides (∆U, GlcNAc). This heparinoid had a lower hemorrhagic effect (400μg/ml) when compared to unfractiionated heparins(25μg/ml).The results also suggest a negligible APTT activity of this heparinoid (62.2s). No action was observed on PT indicating that F-3.0M haven t action on the extrinsic pathway. The results showed that the fraction F- 3.0M have inhibitory effect on migration of leukocytes, 64.5% in the concentration of 10 μg/ml (P<0.001). The search for new heparin and/or heparan sulphates analogs devoid of anticoagulant activity is an atractive alternative and may open up a wide variety of new therapeutic applications

Estudo clínico-laboratorial e histopatológico de cães naturalmente infectados por Leishmania chagasi com diferentes graus de manifestação física

Canine Visceral Leishmania (CVL) is an important zoonotic disease that has a world wide distribution and has a large impact on public health on the American Continent, especially in Brazil, where the nature of endemic diseases in humans affects a large part of the nation. The influence of the prevalence of CVL in the increased rate of human cases in endemic areas and in the unleashing of epidemic outbreaks shows the need for a more profound understanding, that would generate significant advances in the current measures used to control the reservoirs of sickness that are practiced by the Programa Nacional de Vigilância e Controle da Leishmaniose Visceral. The present work describes and compares the clinical-laboratorial and histopathological findings of twenty-three dogs that were naturally infected by Leishmania chagasi, from endemic areas in metropolitan Natal, Rio Grande do Norte, Brazil. These animals, that were selected and given physical and serological exams (IFI and ELISA rK-39), were classified according to the degree of clinical severity and had blood samples drawn (whole blood and serum) for a complete hemogram and a coagulogram to be done as well as biochemical tests for kidney and liver function. The confirmation of infection by L. chagasi was done after the euthanasia of the animals, through the direct demonstration of the parasite in the impression of the spleen and liver crowned with GIEMSA and through a cultivation by means of NNN/Schneider. According to the clinical evaluation, the animals were classified as asymptomatic (7), oligosymptomatic (7) and polysymptomatic (9). Among the animals that were chosen to be autopsied, there were 2 asymptomatic, 3 oligosymptomatic and 3 polysymptomatic, for the purpose of studying their histopathology, having collected fragments of the spleen, liver, kidneys and skin and were fixed in 10% tamponed formol. The comparison between the average parameters of the clinical-laboratory tested animals in the groups was done through the Student t test (a<0.05). The main clinical signals observed were lymphadenomegaly, alopecy, dermatitis, exfoliation, cutaneous ulcers, onicogriphosis and emaciation. The main clinical-laboratorial alterations established, mainly in the polysymptomatic group, were anemia, hyperproteinemia, hyperglobulinemia, alterations in the albumin/globulin ratio and increased ALT activity. Renal alterations were not verified (urea and creatinine levels were normal). Thrombocytopenia was observed in three clinical groups. However, the other indicators of coagulation function (TAP and TTPA) did not have abnormal variations. There were inflammatory infiltrations and leishmania amastigotes in the skin of polysymptomatic dogs, however, they were not found in the skin of asymptomatic animals. Hypertrophy and hyperplasia of the phagocyte mononuclear system, leishmania amastigote parasites were found in the macrophages, extramedullary hematopoiesis and degenerative alterations were detected in the spleen and liver of 8 of the animals submitted to histopathological exams. In accord with these results, it was demonstrated that the expected alterations in the hematological and biochemical parameters in function of their viscerotropic nature of CVL are mainly observed in the more advanced stages of the disease. The absence of inflammatory infiltration and parasite load in the skin suggest that infected animals without symptoms may have an importance irrelevant to the infectiousness of the vector

Extração, caracterização e atividades biológicas de proteínas da espécie cnidoscolus urens (L.) Arthur

The extraction, chemical and structural characterization of a wide variety of compounds derived from plants has been a major source of bioactive molecules. Several proteases have been isolated in the plant kingdom, with numerous pharmacological and biotechnological applications. Among the proteases isolated from plants, are the fibrinogenolytic, with relevant application in the treatment of disorders in the coagulation cascade, in addition to potential use as a tool in clinical laboratories. In this study, in addition to evaluating the effects of the protein extract of Cnidoscolus urens (L.) Arthur (Euphorbiaceae) in the coagulation cascade also investigates the presence of antimicrobial activity and characterizes the proteolytic activity detected in this extract, aiming to determine their potential pharmacological and biotechnological application. In this way, crude protein extracts obtained from the leaves of C. urens in Tris-HCl 0.05M, NaCl 0.15M, pH 7.5, were precipitated in different concentrations of acetone, and assessed for the presence of proteolytic activity in azocaseína and fibrinogen. The most active fraction (F1.0) in these tests was chosen for assessment of biological activity and biochemical characterization. The Aα chain and Bβ of fibrinogen were completely cleaved at a concentration of 0.18 μg/μL of protein fraction in 4 minutes. Fibrinogenolytic activity presented total inhibition in the presence of E-64 and partial in the presence of EDTA. The fraction demonstrated coagulant activity in plasm and reduced the APTT, demonstrating acting on the factors coagulation of the intrinsic pathway and common, not exerting effects on the PT. Fibrinolytic activity on plasma clot was detected only in SDS-PAGE in high concentrations of fraction, and there were no defibrinating. Although several proteases isolated from plants and venomous animals are classically toxic, the fraction F1.0 of C. urens not expressed hemorrhagic nor hemolytic activities. Fraction F1.0 also showed no antimicrobial activity. In proteolytic activity on the azocasein, the optimal pH was 5.0 and optimum temperature of 60ºC. The enzyme activity has been shown to be sensitive to the presence of salts tested, with inhibition for all compounds. The surfactant triton did not influence the enzyme activity, but the tween-20 and SDS inhibited the activity. In the presence of reducing agents increase in enzyme activity occurred, a typical feature of enzymes belonging to the class of cysteine proteases. Several bands with proteolytic activity were detected in zymogram, in the region of high-molecular-weight, which were inhibited by E-64. In this study, we found that C. urens presents in its constitution cysteine proteases with fibrinogenolytic and procoagulant activity, which may be isolated, with potential application in treatment of bleeding disorders, thrombolytic and clinical laboratory

Aplicação da água residuária das salinas no tratamento de efluente têxtil

The treatment of colored and alkaline effluent has been a challenge to the textile industry. An alternative to remove the colors of those effluents is applying magnesium chloride as a coagulant agent. The magnesium ion, in high pH, hydrolyzes itself, forming the magnesium hydroxide which has a large adsorptive area and positive electrostatic charges able to act as an efficient coagulant. The bittern wastewater from the salt industries has been studied as a potential font of this magnesium ion. Nowadays, this bittern wastewater is evicted into the sea, without any treatment or other use. This thesis has evaluated the potential of applying the wastewater from the salt industries in the treatment of dyeing effluent containing indigo dye and alkaline pH. All the experiments were made in jar tests simulating the chemical coagulation, flocculation and decantation steps ranging the pH and the concentration of magnesium ion. Were obtained removals between 96% and 76% for turbidity, apparent color, and true color, respectively, using 200mg/L Mg2+. The reduction of costs with acid, when were used the salt industries wastewater, comparing with Al2(SO4)3, was 62%. For the degradation of organic matter remaining in the clarified, around 900 mg/L, was applyed the advanced process of oxidation: photo-Fenton. The preliminary results showed 57% reduction in DOC. According to the results obtained, the salt industries wastewater can be applied, as coagulant, in the physical-chemical treatment of the denim dyeing wastewater, so it is not necessary a previous adjust of pH, efficiently and economically

Ensaios de tratabilidade aplicados a determinação de parâmetros para dimensionamento de processos de tratamento de água de manancial eutrofizado

Eutrophication is a growing process present in the water sources located in the northeast of Brazil. Among the main consequences of these changes in trophic levels of a water source, stands out adding complexity to the treatment to achieve water standards. By these considerations, this study aimed to define, on a laboratory scale, products and operational conditions to be applied in the processing steps using raw water from Gargalheiras dam, RN, Brazil. The dam mentioned shows a high number of cyanobacteria, with a concentration of cells / ml higher than that established by Decree No. 518/04 MS. The same source was also considered by the state environmental agency in 2009 as hypereutrophic. The static tests developed in this research simulated direct filtration (laboratory filters) and pre-oxidation with chlorine and powdered activated carbon adsorption. The research included the evaluation of the coagulants aluminum hydrochloride (HCA) and alum (SA). The development of the research investigated the conditions for rapid mixing, the dosages of coagulants and pHs of coagulation by the drawing of diagrams. The interference of filtration rate and particle size of filtering means were evaluated as samples and the time of contact were tested with chlorine and activated carbon. By the results of the characterization of the raw water source it was possible to identify the presence of a high pH (7.34). The true color was significant (29 uH) in relation to the apparent color and turbidity (66 uH and 13.60 NTU), reflecting in the measurement of organic matter: MON (8.41 mg.L-1) and Abs254 (0.065 cm-1). The optimization of quick mix set time of 17", the speed gradient of 700 s-1 in the coagulation with HCA and the time of 20" with speed gradient of 800 s-1 for SA. The smaller particle sizes of sand filtering means helped the treatment and the variation in filtration rate did not affect significantly the efficiency of the process. The evaluation of the processing steps found adjustment in standard color and turbidity of the Decree nº 518/04 MS, taking in consideration the average values found in raw water. In the treatment using the HCA for direct filtration the palatable pattern based on the apparent color can be achieved with a dose of 25 mg L-1. With the addition of pre-oxidation step, the standard result was achieved with a reduced dose for 12 mgHCA.L-1. The turbidity standard for water was obtained by direct filtration when the dose exceeds 25 mg L-1 of HCA. With pre-oxidation step there is the possibility of reducing the dose to 20 mg L-1.The addition of CAP adsorption, promoted drinking water for both parameters, with even lower dosage, 13 mg L-1 of HCA. With coagulation using SA removal required for the parameter of apparent color it was achieved with pre-oxidation and 22 mgSA.L-1. Despite the satisfactory results of treatment with the alum, it was not possible to provide water with turbidity less than 1.00 NTU even with the use of all stages of treatment

Avaliação da flotação por ar dissolvido no polimento de efluentes de lagoa de estabilização

Experience in the use of stabilization ponds shows that this is a system with low cost, easy operation and maintenance and suitable for tropical countries. One of its disadvantages is due to high concentrations of suspended solids, mainly due to algal biomass in the effluents. Accordingly, the dissolved air flotation has been shown to be an efficient post-treatment technology. However, the efficiency of this process is related to coagulation and flocculation steps, as well as the adjustment of the involved variables in the process. The objective of this work was to evaluate the algae removal efficiency from wastewater stabilization ponds and the influence of the factors involved in the process using dissolved air flotation. For this, we used primary facultative and maturation effluents of ETE Ponta Negra in Natal. We did tests of coagulation, flocculation and flotation with the samples, using the equipment flotatest. In this process were tested coagulants aluminum sulfate and ferric chloride, and varied the factors pH, coagulant concentration, polymer concentration and rate of recirculation. At the end of the experiments were analyzed turbidity, suspended solids, color, COD and chlorophyll "a". These results were submitted to descriptive statistics to verify the efficiency of the process in general, and regression analysis to identify models that describe the process and demonstrate the factors that have greater influence on flotation. After step methodology, high values were found removal efficiency of suspended solids, reaching values greater than 90% in the best cases. It was concluded that flotation is more efficient in the facultative pond effluent using ferric chloride, and the variability of algae may negatively influence the process. Regression analyzes showed that pH is the most influential variable in the coagulation-flocculation-flotation, and its optimal value among the tested is 5.5 for both coagulants